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. 2019 May 6:6:416-425.
doi: 10.1016/j.toxrep.2019.05.001. eCollection 2019.

Zingiber officinale (Roscoe) mitigates CCl4-induced liver histopathology and biochemical derangements through antioxidant, membrane-stabilizing and tissue-regenerating potentials

Affiliations

Zingiber officinale (Roscoe) mitigates CCl4-induced liver histopathology and biochemical derangements through antioxidant, membrane-stabilizing and tissue-regenerating potentials

Gracious Oluwamayowa Oke et al. Toxicol Rep. .

Abstract

The world-wide increasing incidence of liver injury has attracted scientific interest in the exploration of better treatment or adjuvant treatment therapies. This study investigated the effects of methanol extract of Zingiber officinale (Roscoe) rhizome (MEZOR) in a Wistar rat model of carbon tetrachloride (CCl4)-induced liver injury. The study recruited thirty female Wistar rats that received graded doses of MEZOR (determined by its LD50) by oral gavage through an oral canula, for 4 consecutive weeks following 1 week oral administration of CCl4 (0.7 ml/kg in olive oil; 1:1, v/v) while livolin forte® (5.2 mg/kg p.o.) was used as a standard. CCl4 induced deleterious hepatic effects as revealed by the liver function biomarkers (AST, ALT, ALP and total protein), antioxidant indicators (GSH and CAT) and histopathological effects, demonstrated by H & E, Gordon and Sweet, Masson's trichrome, PAS staining techniques as well as by quantificational analyses of the liver micrographs, using image-J. MEZOR treatment was associated with a dose-dependent and significant mitigation of the aforementioned parameters (p < 0.05). This study concluded that MEZOR is a potential therapeutic choice in the adjuvant treatment of subjects with chemically-induced liver injury.

Keywords: CCl4; Female Wistar rats; Histopathology; Liver injury; Zingiber officinale.

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Figures

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Graphical abstract
Fig. 1
Fig. 1
Effects of MEZOR on [a.] Percentage Weight Change (%), and [b.] Relative Liver Weight (%) of Female Wistar Rats Exposed to CCl4 Toxicity. Each bar represents mean ± standard error of mean at p < 0.05. [1] to [6] = Groups 1–6. * = significant difference when compared with the control group; and a = significant difference when compared with the toxic group.
Fig. 2
Fig. 2
Effects of MEZOR on the Number of Hepatocyte Counts of Female Wistar Rats with CCl4-induced Liver Injury. Each bar represents mean ± standard error of mean at p < 0.05. [1] to [6] = Groups 1–6. * = significant difference when compared with the control group; and a = significant difference when compared with the toxic group.
Fig. 3
Fig. 3
Effects of MEZOR on the Percentage Area of Reticular Fibre (%) of Female Wistar Rats with CCl4-induced Liver Injury. Each bar represents mean ± standard error of mean at p < 0.05. [1] to [6] = Groups 1–6. * = significant difference when compared with the control group. a = significant difference when compared with the toxic group; and b = significant difference when compared with the standard group.
Fig. 4
Fig. 4
Effects of MEZOR on the Percentage Area of Collagen Fibre (%) of Female Wistar Rats with CCl4-induced Liver Injury. Each bar represents mean ± standard error of mean at p < 0.05. [1] to [6] = Groups 1–6. * = significant difference when compared with the control group. a = significant difference when compared with the toxic group; and b = significant difference when compared with the standard group.
Fig. 5
Fig. 5
Effects of MEZOR on Total Protein Level in Female Wistar Rats with CCl4-induced Liver Injury. Each bar represents mean ± standard error of mean at p < 0.05. [1] to [6] = Groups 1–6. * = significant difference when compared with the control group. a = significant difference when compared with the toxic group; and b = significant difference when compared with the standard group.
Fig. 6
Fig. 6
Effects of MEZOR on [a.] GSH Level, and [b.] CAT Activity in Female Wistar Rats with CCl4-induced Liver Injury. Each bar represents mean ± standard error of mean at p < 0.05. [1] to [6] = Groups 1–6. * = significant difference when compared with the control group. a = significant difference when compared with the toxic group; and b = significant difference when compared with the standard group.
Fig. 7
Fig. 7
Histological Effects of MEZOR in Female Wistar Rats with CCl4-induced Liver Injury; General Histoarchitecture Using H & E Staining Technique. Fig. 7a Light micrographs of the control and toxic groups showing the centrilobular zones Scale bar =50 μm. Fig. 7b Light micrographs of the standard (Livolin) and MEZOR-treated groups showing the centrilobular zones. Scale bar =50 μm. Fig. 7c Light micrographs of the standard (Livolin) and MEZOR-treated groups showing the intermediate zones. Scale bar =50 μm. *1 and 2 = Groups 1 and 2. Plates 1a and 2a = intermediate zones of liver histoarchitecture. Plates 1b and 2b = portal areas of liver histoarchitecture. *3 to 6 = Groups 3–6. Scale bar =50 μm. Long black arrow = apparently normal hepatocytes; CV = centrilobular vein; black dashed arrow = hepatic sinusoid; short red arrow = necrotic hepatocytes with nuclei karyolysis; short orange arrow = necrotic hepatocyte with heperchromatic nuclei; short green arrow = necrotic hepatocyte with fragmented nuclei; short blue arrow = massive proliferation of Kupffer cells at the luminal surface of the hepatic sinusoids; short black arrow = intranuclear vacuolation; long blue arrow = hepatic hepatocyte with intracytoplasmic vacuolation; black dotted circle = portal space; long yellow arrow = infiltration of the centrilobular zone by inflammatory cells; blach dotted arc = area of discontinuation of the simple squamous epithelial lining of the central vein; short blue arrow = massive proliferation of kupffer cells at the luminal surface of the hepatic sinusoids; green asterisk = artifact.
Fig. 8
Fig. 8
Histological effects of MEZOR on reticular fibre formation in Female Wistar Rats with CCl4-induced Liver Injury. Gordon and Sweet’s silver stain (Magnification x 400; Scale bar =50 μm). * [1] to [6] = Groups 1–6. Long black arrow = apparently normal hepatocyte; yellow arrow = dark stained apparently intact reticular fibres surrounding individual hepatocyte; deep blue arrow = reticular fibre spanning across the perisinusoidal space; red arrow = reticular fibre spanning across the wall of the centrilobular vein; short black arrow = discontinuous reticular fibres surrounding the centrilobular vein; red dotted circle and light blue arrow = intracytoplasmic vacuolation; CV = centrilobular vein.
Fig. 9
Fig. 9
a Histological effects of MEZOR on collagen fibre formation in Female Wistar Rats with CCl4-induced Liver Injury. Masson’s Trichrome Staining Technique (Magnification x100; Scale bar =200 μm). * [1] to [6] = Groups 1–6. Dark blue dotted circle = portal space; short dark red arrow = deposits of light green-stained collagen fibres. Fig. 9b Histological effects of MEZOR on collagen fibre formation in the hepatic centrilobular zone of Female Wistar Rats with CCl4-induced Liver Injury. Masson’s Trichrome Staining Technique (Magnification x400; Scale bar =50 μm). * [1] to [6] = Groups 1–6. CV = centrilobular vein; short black arrow = light green-stained collagen fibres; short black arrow = dilation of centrilobular vein with marked deposits of collagen fibres; dotted circle = parenchymal tissue of the centrilobular zone.
Fig. 10
Fig. 10
Histological effects of MEZOR on the hepatic glycogen content of Female Wistar Rats with CCl4-induced Liver Injury. Period Acid-Schiff (PAS) stain (magnification = ×400; Scale bar =50 μm). * [1] to [6] = Groups 1–6. Black arrow = intracytoplasmic PAS-positive substance (glycogen); CV = centrilobular vein; red arrow = intracytoplasmic vacuolation; insets = light micrographs with PAS stain and diastase (PAS + D) showing digestion of hepatic lobule glycogen content by diastase.

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